Skip to main content

Genomic Inverse PCR for Screening of Preleukemic Cells in Newborns (GIPFEL Technology)

Part of the Methods in Molecular Biology book series (MIMB,volume 2185)

Abstract

Recurrent chromosomal translocations define genetic subtypes of childhood leukemia and present the first hit that generates an expanded clone of preleukemic cells in the bone marrow. Most commonly, reverse transcriptase PCR is used to detect these translocations on RNA level. This technique has severe drawbacks, including sensitivity to contamination and instability of RNA. Here, we describe the genomic inverse PCR for exploration of ligated breakpoints (GIPFEL) that overcomes these pitfalls.

Key words

  • GIPFEL
  • Chromosomal translocation
  • Newborn screening
  • Preleukemic cells

This is a preview of subscription content, access via your institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • DOI: 10.1007/978-1-0716-0810-4_8
  • Chapter length: 22 pages
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
eBook
USD   119.00
Price excludes VAT (USA)
  • ISBN: 978-1-0716-0810-4
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
Softcover Book
USD   159.99
Price excludes VAT (USA)
Hardcover Book
USD   219.99
Price excludes VAT (USA)
Fig. 1
Fig. 2

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Mori H, Colman SM, Xiao Z, Ford AM, Healy LE, Donaldson C, Hows JM, Navarrete C, Greaves M (2002) Chromosome translocations and covert leukemic clones are generated during normal fetal development. Proc Natl Acad Sci U S A 99(12):8242–8247. https://doi.org/10.1073/pnas.112218799

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  2. Schäfer D, Olsen M, Lähnemann D, Stanulla M, Slany R, Schmiegelow K, Borkhardt A, Fischer U (2018) Five percent of healthy newborns have an ETV6-RUNX1 fusion as revealed by DNA-based GIPFEL screening. Blood 131(7):821–826. https://doi.org/10.1182/blood-2017-09-808402

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  3. Ford AM, Ridge SA, Cabrera ME, Mahmoud H, Steel CM, Chan LC, Greaves M (1993) In utero rearrangements in the trithorax-related oncogene in infant leukaemias. Nature 363(6427):358–360. https://doi.org/10.1038/363358a0

  4. Gill Super HJ, Rothberg PG, Kobayashi H, Freeman AI, Diaz MO, Rowley JD (1994) Clonal, nonconstitutional rearrangements of the MLL gene in infant twins with acute lymphoblastic leukemia: in utero chromosome rearrangement of 11q23. Blood 83(3):641–644

    Google Scholar 

  5. Megonigal MD, Rappaport EF, Jones DH, Williams TM, Lovett BD, Kelly KM, Lerou PH, Moulton T, Budarf ML, Felix CA (1998) T(11;22)(q23;q11.2) in acute myeloid leukemia of infant twins fuses MLL with hCDCrel, a cell division cycle gene in the genomic region of deletion in DiGeorge and velocardiofacial syndromes. Proc Natl Acad Sci U S A 95(11):6413–6418. https://doi.org/10.1073/pnas.95.11.6413

  6. Hein D, Dreisig K, Metzler M, Izraeli S, Schmiegelow K, Borkhardt A, Fischer U (2019) The preleukemic TCF3-PBX1 gene fusion can be generated in utero and is present in approximately 0.6% of healthy newborns. Blood 134(16):1355–1358. https://doi.org/10.1182/blood.2019002215

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  7. Pui CH, Carroll WL, Meshinchi S, Arceci RJ (2011) Biology, risk stratification, and therapy of pediatric acute leukemias: an update. J Clin Oncol 29(5):551–565. https://doi.org/10.1200/jco.2010.30.7405

    CrossRef  PubMed  Google Scholar 

  8. Shibata Y, Malhotra A, Dutta A (2010) Detection of DNA fusion junctions for BCR-ABL translocations by anchored ChromPET. Genome Med 2(9):70. https://doi.org/10.1186/gm191

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  9. Füller E, Schäfer D, Fischer U, Krell PF, Stanulla M, Borkhardt A, Slany RK (2014) Genomic inverse PCR for exploration of ligated breakpoints (GIPFEL), a new method to detect translocations in leukemia. PLoS One 9(8):e104419. https://doi.org/10.1371/journal.pone.0104419

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Daniel Hein .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and Permissions

Copyright information

© 2021 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Verify currency and authenticity via CrossMark

Cite this protocol

Hein, D., Borkhardt, A., Fischer, U. (2021). Genomic Inverse PCR for Screening of Preleukemic Cells in Newborns (GIPFEL Technology). In: Cobaleda, C., Sánchez-García, I. (eds) Leukemia Stem Cells. Methods in Molecular Biology, vol 2185. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0810-4_8

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-0810-4_8

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0809-8

  • Online ISBN: 978-1-0716-0810-4

  • eBook Packages: Springer Protocols